Cell-type specific changes in PKC-delta neurons of the central amygdala during alcohol withdrawal

Dilly, Geoffrey A.; Kittleman, Cory W.; Kerr, Tony M.; Messing, Robert O.; Mayfield, R. Dayne

doi:10.1038/s41398-022-02063-0

Cited by 29 publications

(18 citation statements)

References 67 publications

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“…This uncovered many new CEA cell types, their participation in CEA circuitry, and revealed their relationships to previously identified CEA marker genes. Recently, single-nucleus RNA sequencing from the CEA in rat identified 13 neuronal types (11/13 neuronal types were from CEA) ( Dilly et al, 2022 ). We found good correspondence with major CEA cell types identified by Dilly et al, such as the Prkcd and the Crh neurons in the CeC and CeL, and the Drd1 and Drd2 neurons in the CeM.…”

Section: Discussionmentioning

confidence: 99%

Multimodal mapping of cell types and projections in the central nucleus of the amygdala

Wang

Krabbe

Eddison

et al. 2023

eLife

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The central nucleus of the amygdala (CEA) is a brain region that integrates external and internal sensory information and executes innate and adaptive behaviors through distinct output pathways. Despite its complex functions, the diversity of molecularly defined neuronal types in the CEA and their contributions to major axonal projection targets have not been examined systematically. Here, we performed single-cell RNA-sequencing (scRNA-Seq) to classify molecularly defined cell types in the CEA and identified marker genes to map the location of these neuronal types using expansion asisted iterative fluorescence in situ hybridization (EASI-FISH). We developed new methods to integrate EASI-FISH with 5-plex retrograde axonal labeling to determine the spatial, morphological, and connectivity properties of ~30,000 molecularly defined CEA neurons. Our study revealed spatio-molecular organization of the CEA, with medial and lateral CEA associated with distinct molecularly defined cell families. We also found a long-range axon projection network from the CEA, where target regions receive inputs from multiple molecularly defined cell types. Axon collateralization was found primarily among projections to hindbrain targets, which are distinct from forebrain projections. This resource reports marker gene combinations for molecularly defined cell types and axon-projection types, which will be useful for selective interrogation of these neuronal populations to study their contributions to the diverse functions of the CEA.

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Section: Discussionmentioning

confidence: 99%

Multimodal mapping of cell types and projections in the central nucleus of the amygdala

Wang

Krabbe

Eddison

et al. 2023

eLife

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“…In the CeL, we found two clusters marked by expression of Sst: the CeL Sst cluster which contains the largest and most highly expressing population of Sst-positive neurons, and the CeL NTS/Tac2 cluster which contains low expressing Sst-positive cells. This is consistent with previous observations (Kim et al ., 2017; McCullough et al ., 2018) and a recent single cell transcriptomics study in the rat CeA (Dilly et al, 2022). Optogenetic manipulation of these two clusters separately suggested that both drive appetitive behavior (Kim et al ., 2017).…”

Section: Discussionmentioning

confidence: 99%

Transcriptomics reveals amygdala neuron regulation by fasting and ghrelin thereby promoting feeding

Peters

Fermani

et al. 2022

Preprint

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SUMMARYThe central amygdala (CeA) consists of numerous genetically-defined inhibitory neurons that control defensive and appetitive behaviors including feeding. Transcriptomic signatures of cell types and their links to function remain poorly understood. Using single-nucleus RNA-sequencing, we describe nine CeA cell clusters, of which four are mostly associated with appetitive and two with aversive behaviors. To analyze the activation mechanism of appetitive CeA neurons, we characterized Serotonin receptor 2a (Htr2a)-expressing neurons (CeAHtr2a) that comprise three appetitive clusters and were previously shown to promote feeding. In vivo calcium imaging revealed that CeAHtr2aneurons are activated by fasting, the hormone ghrelin, and the presence of food. Moreover, these neurons are required for the orexigenic effects of ghrelin. Appetitive CeA neurons responsive to fasting and ghrelin project to the parabrachial nucleus (PBN) causing inhibition of target PBN neurons. These results illustrate how the transcriptomic diversification of CeA neurons relates to fasting and hormone-regulated feeding behavior.HighlightsTranscriptomics reveals four appetitive, two aversive and three unknown CeA clustersGhrelin and fasting activate CeAHtr2aneurons comprising three appetitive clustersGhrelin’s orexigenic effects require the activity of appetitive CeA neuronsCeA-→PBN inhibition is necessary for ghrelin/fasting induced feeding

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“…This transcriptome signature is less common in mammals (e.g., refs. 59 , 68 , 69 ). We did not detect other neuron types in this brain region.…”

Section: Resultsmentioning

confidence: 99%

A custom-made AAV1 variant (AAV1-T593K) enables efficient transduction of Japanese quail neurons in vitro and in vivo

et al. 2023

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The widespread use of rodents in neuroscience has prompted the development of optimized viral variants for transduction of brain cells, in vivo. However, many of the viruses developed are less efficient in other model organisms, with birds being among the most resistant to transduction by current viral tools. Resultantly, the use of genetically-encoded tools and methods in avian species is markedly lower than in rodents; likely holding the field back. We sought to bridge this gap by developing custom viruses towards the transduction of brain cells of the Japanese quail. We first develop a protocol for culturing primary neurons and glia from quail embryos, followed by characterization of cultures via immunostaining, single cell mRNA sequencing, patch clamp electrophysiology and calcium imaging. We then leveraged the cultures for the rapid screening of various viruses, only to find that all yielded poor to no infection of cells in vitro. However, few infected neurons were obtained by AAV1 and AAV2. Scrutiny of the sequence of the AAV receptor found in quails led us to rationally design a custom-made AAV variant (AAV1-T593K; AAV1*) that exhibits improved transduction efficiencies in vitro and in vivo (14- and five-fold, respectively). Together, we present unique culturing method, transcriptomic profiles of quail’s brain cells and a custom-tailored AAV1 for transduction of quail neurons in vitro and in vivo.

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Cell-type specific changes in PKC-delta neurons of the central amygdala during alcohol withdrawal

Cited by 29 publications

References 67 publications

Multimodal mapping of cell types and projections in the central nucleus of the amygdala

Multimodal mapping of cell types and projections in the central nucleus of the amygdala

Transcriptomics reveals amygdala neuron regulation by fasting and ghrelin thereby promoting feeding

A custom-made AAV1 variant (AAV1-T593K) enables efficient transduction of Japanese quail neurons in vitro and in vivo

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